The present invention concerns secreted factors encoded by genes differentially regulated in certain diseased tissues. More particularly, the invention concerns nucleic acid encoding novel secreted polypeptide factors, the encoded polypeptides, and compositions containing and methods and means for producing them. The invention further concerns methods based on the use of such nucleic acids and/or polypeptides in the diagnosis and treatment of various diseases, in particular cardiac, renal, or inflammatory diseases.
Gene expression patterns, including changes in gene expression between normal and diseased tissues or tissues in various stages of disease progression provide valuable insight into the molecular determinants of normal and abnormal cellular physiology. Accordingly, genes that are differentially expressed in subjects suffering from a disease, such as cardiac, renal or inflammatory disease, relative to normal subjects, are useful targets for intervention to diagnose, prevent or treat such diseases.
Techniques have been developed to efficiently analyze the level of expression of specific genes in cells and tissues. Procedures that can be used to identify and clone differentially expressed genes include, for example, subtractive hybridization (Jiang and Fisher, Mol. Cell. Different. 1:285-299 [1993]; Jiang etal., Oncogene 10, 1855-1864 [1995]; Sagerstrom et al., Annu. Rev. Biochem. 66: 751-783 [1997]); differential RNA display (DDRT-PCR) (Watson et al., Developmental Neuroscience 15:77-86 [1993]; Liang and Pardee, Science 257:967-971 [1992]); RNA fingerprinting by arbitrarily primed PCR (RAP-PCR) (Ralph et al., Proc. Natl. Acad. Sci. USA 90:10710-10714 [1993]; McClelland and Welsh, PCR Methods and Applications 4:S66-81 [1994]); representational difference analysis (RDA) (Hubank and Schatz, Nucl. Acids Res. 22:5640-5648 [1994]); serial analysis of gene expression (SAGE) (Velculescu et al., Science 270:484-487 [1995]; Zhang et al., Science 276:1268-1272 [1997]); electronic subtraction (Wan et al., Nature Biotechnology 4:1685-1691 [1996]); combinatorial gene matrix analyses (Schena et al., Science 270:467-470 [1995]), and various modifications and improvements of these and similar techniques.
A particularly attractive method for assessing gene expression is the DNA microarray technique. In this method, nucleotide sequences of interest are plated, or arrayed, on a porous or non-porous substrate that can be paper, nylon or other type of membrane, filter, chip, glass slide or any other suitable solid support. The arrayed sequences are then hybridized with specific DNA probes from cells or tissues of interest. Microarrays of biological materials have been described in a number of patents and patent applications, including, for example, U.S. Pat. Nos. 5,744,305; 5,800,992; 5,807,522; 5,716,785; and European Patent No. 0 373 203.
The DNA microarray technique can be used to monitor the expression level of large numbers of genes simultaneously (to produce a transcript image), and to identify genetic variants, mutations and polymorphisms. This information may be used to determine gene function, understanding the genetic basis of disease, diagnosing disease, and developing and monitoring the activities of therapeutic agents.
An important application of the microarray method allows for the assessment of differential gene expression in pairs of mRNA samples from two different tissues, or in the same tissue comparing normal versus disease states or time progression of the disease. Microarray analysis allows one to analyze the expression of known genes of interest, or to discover novel genes expressed differentially in tissues of interest. Thus, an attractive application of this technology is as a fundamental discovery tool to identify new genes, and their corresponding expression products, which contribute to the pathogenesis of disease and related conditions.
Microarray technology has been successfully applied to large-scale analysis of human gene expression to identify cancer-specific genes and inflammatory-specific genes (DeRisi et al., Nat. Genet., 14(4):457-60 [1996]; Heller et al., Proc. Natl. Acad. Sci. USA, 94(6):2150-55 [1997]). DeRisi et al. examined a pre-selected set of 870 different genes for their expression in a melanoma cell line and a non-tumorigenic version of the same cell line. The microarray analysis revealed a decrease in expression for 15/870 (1.7%) and an increase in expression for 63/870 (7.3%) of the genes in non-tumorigenic relative to tumorigenic cells (differential expression values  less than 0.52 or  greater than 2.4 were deemed significant). Heller et al. employed microarrays to evaluate the expression of 1000 genes in cells taken from normal and inflamed human tissues. The results indicated that altered expression was evident in genes encoding inflammatory mediators such as IL-3, and a tissue metalloprotease. These results illustrate the utility of applying microarray technology to complex human diseases.
It would be beneficial to discover differentially expressed genes that are related to diseases or various disease states. It would further be beneficial to develop methods and compositions for the diagnostic evaluation and prognosis of conditions involving such diseases, for the identification of subjects exhibiting a predisposition to such conditions, for modulating the effect of these differentially expressed genes and their expression products, for monitoring patients undergoing clinical evaluation for the prevention and treatment of a disease, specifically cardiac, kidney or inflammatory disease, and for monitoring the efficacy of compounds used in clinical trials.
Secreted proteins mediate key biological processes including cell to cell interactions as well as important cellular functions such as cell growth and differentiation, and most protein-based drugs are secreted proteins including insulin, growth hormone, interferons, tissue plasminogen activator ( tPA), and erythropoietin (EPO). It would, therefore, be particularly desirable to identify novel differentially expressed genes encoding secreted proteins.
In one aspect, the present invention concerns an isolated nucleic acid molecule comprising a poly- or oligonucleotide selected from the group consisting of:
(a) a polynucleotide encoding a polypeptide having at least about 80% sequence identity with amino acids selected from the group consisting of: 1 to 1203 of SEQ ID NO: 2, amino acids 1 to 193 of SEQ ID NO: 4, amino acids 1 to 236 of SEQ ID NO:6, amino acids 1 to 61 of SEQ ID NO: 8, amino acids 1 to 79 of SEQ ID NO:10, amino acids 1 to 92 of SEQ ID NO:12, amino acids 1 to 86 of SEQ ID NO:14, amino acids 1 to 36 of SEQ ID NO:16, amino acids 1 to 83 of SEQ ID NO:18, amino acids 1 to 82 of SEQ ID NO:20, amino acids 1 to 462 of SEQ ID NO:22, amino acids 1 to 170 of SEQ ID NO:24, amino acids xe2x88x9226 to 233 of FIG. 13 (amino acids 1 to 259 of SEQ ID NO:26), amino acids 1 to 30 of SEQ ID NO:28, amino acids 1 to 39 of SEQ ID NO:30, amino acids 1 to 541 of SEQ ID NO: 33, amino acids 1 to 30 of SEQ ID NO:35, amino acids 1 to 100 of SEQ ID NO:37, amino acids 1 to 65 of SEQ ID NO:39, amino acids 1 to 42 of SEQ ID NO:41, amino acids 1 to 46 of SEQ ID NO:43, amino acids 1 to 313 of SEQ ID NO:46, amino acids 1 to 58 of SEQ ID NO:51, amino acids xe2x88x9235 to 387 of FIG. 29 (amino acids 1 to 422 of SEQ ID NO:53), amino acids 1 to 58 of SEQ ID NO:55, amino acids 1 to 52 of SEQ ID NO:57, amino acids 1 to 245 of SEQ ID NO:59, amino acids 1 to 142 of SEQ ID NO:63, amino acids 1 to 49 of SEQ ID NO:67, amino acids 1 to 70 of SEQ ID NO:69, amino acids 1 to 113 of SEQ ID NO: 72, and amino acids 1 to 114 of SEQ ID NO:74, and amino acids 1 to 97 of SEQ ID NO:76; or a transmembrane domain (membrane spanning segment/region) deleted or inactivated variant thereof;
(b) a polynucleotide encoding a polypeptide having at least about 80% sequence identity with amino acids 1 to 233 of SEQ ID NO: 26, or amino acids 1 to 387 of SEQ ID NO: 53;
(c) a polynucleotide encoding amino acids selected from the group consisting of: 1 to 203 of SEQ ID NO: 2, amino acids 1 to 193 of SEQ ID NO: 4, amino acids 1 to 236 of SEQ ID NO:6, amino acids 1 to 61 of SEQ ID NO: 8, amino acids 1 to 79 of SEQ ID NO:10, amino acids 1 to 92 of SEQ ID NO:12, amino acids 1 to 86 of SEQ ID NO:14, amino acids 1 to 36 of SEQ ID NO:16, amino acids 1 to 83 of SEQ ID NO:18, amino acids 1 to 82 of SEQ ID NO:20, amino acids 1 to 462 of SEQ ID NO:22, amino acids 1 to 170 of SEQ ID NO:24, amino acids xe2x88x9226 to 233 of FIG. 13 (amino acids 1 to 259 of SEQ ID NO:26), amino acids 1 to 30 of SEQ ID NO:28, amino acids 1 to 39 of SEQ ID NO:30, amino acids 1 to 541 of SEQ ID NO: 33, amino acids 1 to 30 of SEQ ID NO:35, amino acids 1 to 100 of SEQ ID NO:37, amino acids 1 to 65 of SEQ ID NO:39, amino acids 1 to 42 of SEQ ID NO:41, amino acids 1 to 46 of SEQ ID NO:43, amino acids 1 to 313 of SEQ ID NO:46, amino acids 1 to 58 of SEQ ID NO:51, amino acids xe2x88x9235 to 387 of FIG. 29 (amino acids 1 to 422 of SEQ ID NO:53), amino acids 1 to 58 of SEQ ID NO:55, amino acids 1 to 52 of SEQ ID NO:57, amino acids 1 to 245 of SEQ ID NO:59, amino acids 1 to 142 of SEQ ID NO:63, amino acids 1 to 49 of SEQ ID NO:67, amino acids 1 to 70 of SEQ ID NO:69, amino acids 1 to 113 of SEQ ID NO: 72, and amino acids 1 to 114 of SEQ ID NO:74, and amino acids 1 to 97 of SEQ ID NO:76; or a transmembrane domain (membrane spanning segment/region) deleted or inactivated variant thereof;
(d) a polynucleotide selected from the group consisting of: a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 1, wherein said polynucleotide encodes a polypeptide having at least one biological activity o [the polypeptide encoded by clone P00184_D11 (SEQ ID NO: 1), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 3, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00185_D11 (SEQ ID NO: 3); a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 5, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00188_D12 (SEQ ID NO: 5), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 7, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00188_E01 (SEQ ID NO: 7), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 9, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00194_G01 (SEQ ID NO: 9), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 11, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00194_G05 (SEQ ID NO: 11), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 13, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00194_H10 (SEQ ID NO:13), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 15, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00199_D08 (SEQ ID NO: 15), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 17, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00203_D04 (SEQ ID NO: 17), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 19, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00203_E06 (SEQ ID NO: 19), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 21, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00209_F06 (SEQ ID NO: 21), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 23, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00219_D02 (SEQ ID NO: 23), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 25, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00219_F06 (SEQ ID NO: 25), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 27, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00220_H05 (SEQ ID NO: 27), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 29, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00222_G03 (SEQ ID NO: 29), a polynucleotide hybridizing under stringent conditions with the complement of the polynucleotide of SEQ ID NO: 31 (clone P00223_F07), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 32, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00225_C01 (SEQ ID NO: 32), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 34, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00227_D11 (SEQ ID NO: 34), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 36, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00228_F03 (SEQ ID NO: 36), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 38, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00233_H08 (SEQ ID NO: 38), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 40, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00235_G08 (SEQ ID NO: 40), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 42, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00239_C11 (SEQ ID NO: 42), a polynucleotide hybridizing under stringent conditions with the complement of the polynucleotide of SEQ ID NO: 44 (clone P00240_B04), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 45, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00240_E05 (SEQ ID NO: 45), a polynucleotide hybridizing under stringent conditions with the complement of the polynucleotide of SEQ ID NO: 47 (clone P00241_E12), a polynucleotide hybridizing under stringent conditions with the complement of the polynucleotide of SEQ ID NO: 48 (clone P00245_D06), a polynucleotide hybridizing under stringent conditions with the complement of the polynucleotide of SEQ ID NO: 49 (clone P00246_D12), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 50, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00247_A04 (SEQ ID NO: 50), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 52, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00248_B04 (SEQ ID NO: 52), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 54, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00249_F09 (SEQ ID NO: 54), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 56, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00258_A10 (SEQ ID NO: 56), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 58, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00262_C10 (SEQ ID NO: 58), a polynucleotide hybridizing under stringent conditions with the complement of the polynucleotide of SEQ ID NO: 60 (clone P00263_G06), a polynucleotide hybridizing under stringent conditions with the complement of the polynucleotide of SEQ ID NO: 61 (clone P00267_F08) , a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 62, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00269_H08 (SEQ ID NO: 62), a polynucleotide hybridizing under stringent conditions with the complement of the polynucleotide of SEQ ID NO: 64 (clone P00312_C04), a polynucleotide hybridizing under stringent conditions with the complement of the polynucleotide of SEQ ID NO: 65 (clone P00324_H02), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 66, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00628_H02 (SEQ ID NO: 66), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 68, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00629_C08 (SEQ ID NO: 68), a polynucleotide hybridizing under stringent conditions with the complement of the polynucleotide of SEQ ID NO: 70 (clone P00634_G11), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 71, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00641_G11 (SEQ ID NO: 71), a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 73, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00648_E12 (SEQ ID NO: 73), and a polynucleotide hybridizing under stringent conditions with the complement of the coding region of SEQ ID NO: 75 wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00697_C03 (SEQ ID NO: 75);
(e) a polynucleotide encoding at least about 50 contiguous amino acids from amino acids selected from the group consisting of: amino acids 1 to 203 of SEQ ID NO: 2, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00184_D11 (SEQ ID NO: 1), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 193 of SEQ ID NO: 4, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00185_D11 (SEQ ID NO: 3); a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 236 of SEQ ID NO: 6, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00188_D12 (SEQ ID NO: 5), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 61 of SEQ ID NO: 8, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00188_E01 (SEQ ID NO: 7), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 79 of SEQ ID NO: 10, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00194_G01 (SEQ ID NO: 9), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 92 of SEQ ID NO: 12, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00194_G05 (SEQ ID NO: 11), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 86 of SEQ ID NO: 14, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00194_H10 (SEQ ID NO:13), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 36 of SEQ ID NO: 16, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00199_D08 (SEQ ID NO: 15), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 83 of SEQ ID NO: 18, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00203_D04 (SEQ ID NO: 17), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 82 of SEQ ID NO: 20, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00203_E06 (SEQ ID NO: 19), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 462 of SEQ ID NO: 22, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00209_F06 (SEQ ID NO: 21), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 170 of SEQ ID NO: 24, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00219_D02 (SEQ ID NO: 23), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids xe2x88x9226 to 233 of FIG. 13 (amino acids 1 to 259 of SEQ ID NO:26), wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00219_F06 (SEQ ID NO: 25), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 30 of SEQ ID NO: 28, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00220_H05 (SEQ ID NO: 27), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 39 of SEQ ID NO: 30, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00222_G03 (SEQ ID NO: 29), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 541 of SEQ ID NO: 33, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00225_C01 (SEQ ID NO: 32), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 30 of SEQ ID NO: 35, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00227_D11 (SEQ ID NO: 34), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 100 of SEQ ID NO: 37, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00228_F03 (SEQ ID NO: 36), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 65 of SEQ ID NO: 39, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00233_H08 (SEQ ID NO: 38), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 65 of SEQ ID NO: 39, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00235_G08 (SEQ ID NO: 40), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 46 of SEQ ID NO: 43, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00239_C11 (SEQ ID NO: 42), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 313 of SEQ ID NO: 46, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00240_E05 (SEQ ID NO: 45), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 58 of SEQ ID NO: 51, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00247_A04 (SEQ ID NO: 50), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids xe2x88x9235 to 387 of FIG. 29 (amino acids 1 to 422 of SEQ ID NO:53), wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00248_B04 (SEQ ID NO: 52), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 58 of SEQ ID NO: 55, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00249_F09 (SEQ ID NO: 54), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 52 of SEQ ID NO: 57, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00258_A10 (SEQ ID NO: 56), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 245 of SEQ ID NO: 59, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00262_C10 (SEQ ID NO: 58), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 142 of SEQ ID NO: 63, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00269_H08 (SEQ ID NO: 62), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 49 of SEQ ID NO: 67, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00628_H02 (SEQ ID NO: 66), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 70 of SEQ ID NO: 69, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00629_C08 (SEQ ID NO: 68), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 113 of SEQ ID NO: 72, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00641_G11 (SEQ ID NO: 71), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 114 of SEQ ID NO: 74, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00648_E12 (SEQ ID NO: 73), a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 97 of SEQ ID NO: 76, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00697_C03 (SEQ ID NO: 75);
(f) a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 233 of SEQ ID NO: 26, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00219_F06 (SEQ ID NO: 25), and a polynucleotide encoding at least about 50 contiguous amino acids from amino acids 1 to 387 of SEQ ID NO: 53, wherein said polynucleotide encodes a polypeptide having at least one biological activity of the polypeptide encoded by clone P00248_B04 (SEQ ID NO: 52);
(g) a polynucleotide selected from the group consisting of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 32, 34, 36, 38, 40, 42, 44, 45, 47, 48, 49, 50, 52, 54, 56, 58, 60, 61, 62, 64, 65, 66, 68, 70, 71, 73, and 75;
(h) the complement of a polynucleotide of (a)-(g); and
(i) an antisense oligonucleotide capable of hybridizing with, and inhibiting the translation of, the rnRNA encoded by a gene encoding a polypeptide selected from the group consisting of: SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57, 59, 63, 67, 69, 72, 74, 76, and another mammalian (e.g. human) homologue thereof.
In another aspect, the invention concerns a vector comprising any of the poly- or oligonucleotides of (a)-(i) above.
In a further aspect, the invention concerns a recombinant host cell transformed with nucleic acid comprising any of the poly- or oligonucleotides of (a)-(i) above, or with a vector comprising any of the poly- or oligonucleotides of (a)-(i) above.
In a still further aspect, the invention concerns a recombinant method for producing a polypeptide by culturing a recombinant host cell transformed with nucleic acid comprising any of the polynucleotides of (a)-(g) above under conditions such that the polypeptide is expressed, and isolating the polypeptide.
In a different aspect, the invention concerns a polypeptide comprising:
(a) a polypeptide having at least about 80% identity with amino acids 1 to 203 of SEQ ID NO: 2, amino acids 1 to 193 of SEQ ID NO: 4, amino acids 1 to 236 of SEQ ID NO:6, amino acids 1 to 61 of SEQ ID NO: 8, amino acids 1 to 79 of SEQ ID NO:10, amino acids 1 to 92 of SEQ ID NO:12, amino acids 1 to 86 of SEQ ID NO:14, amino acids 1 to 36 of SEQ ID NO:16, amino acids 1 to 83 of SEQ ID NO:18, amino acids 1 to 82 of SEQ ID NO:20, amino acids 1 to 462 of SEQ ID NO:22, amino acids 1 to 170 of SEQ ID NO:24, amino acids xe2x88x9226 to 233 of FIG. 13 (amino acids 1 to 259 of SEQ ID NO:26), amino acids 1 to 30 of SEQ ID NO:28, amino acids 1 to 39 of SEQ ID NO:30, amino acids 1 to 541 of SEQ ID NO:33, amino acids 1 to 30 of SEQ ID NO: 35, amino acids 1 to 100 of SEQ ID NO:37, amino acids 1 to 65 of SEQ ID NO:39, amino acids 1 to 42 of SEQ ID NO:41, amino acids 1 to 46 of SEQ ID NO:43, amino acids 1 to 313 of SEQ ID NO:46, amino acids 1 to 58 of SEQ ID NO:51, amino acids xe2x88x9235 to 387 of FIG. 29 (amino acids 1 to 422 of SEQ ID NO:53), amino acids 1 to 58 of SEQ ID NO:55, amino acids 1 to 52 of SEQ ID NO:57, amino acids 1 to 245 of SEQ ID NO:59, amino acids 1 to 142 of SEQ ID NO:63, amino acids 1 to 49 of SEQ ID NO:67, amino acids 1 to 70 of SEQ ID NO:69, amino acids 1 to 113 of SEQ ID NO:72, amino acids 1 to 114 of SEQ ID NO:74, amino acids 1 to 97 of SEQ ID NO:76; or
a polypeptide encoded by nucleic acid hybridizing under stringent conditions with the complement of the coding region of SEQ ID NOS:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 32, 34, 36, 38, 40, 42, 44, 45, 47, 48, 49, 50, 52 54, 56, 58, 60, 61, 62, 64, 65, 66, 68, 70, 71, 73, 75;
the polypeptides of (a) and (b) having at least one biological activity of the polypeptide encoded by clones P00184_D11 (SEQ ID NO:1), P00185_D11 (SEQ ID NO:3), P00188_D12 (SEQ ID NO:5), P00188_E01 (SEQ ID NO:7), P00194_G01 (SEQ ID NO:9), P00194_G05 (SEQ ID NO:11), P00194_H10 (SEQ ID NO:13), P00199_D08 (SEQ ID NO:15), P00203_D04 (SEQ ID NO:17), P00203_E06 (SEQ ID NO:19), P00209_F06 (SEQ ID NO:21), P00219_D02 (SEQ ID NO:23), P00219_F06 (SEQ ID NO:25), P00220_H05 (SEQ ID NO:27), P00222_G03 (SEQ ID NO:29), P00223_F07 (SEQ ID NO:31), P00225_C01 (SEQ ID NO:32), P00227_D11 (SEQ ID NO:34), P00228_F03 (SEQ ID NO:36), P00233_H08 (SEQ ID NO:38), P00235_G08 (SEQ ID NO:40), P00239_C11 (SEQ ID NO:42), P00240_B04 (SEQ ID NO:44), P00240_E05 (SEQ ID NO:45), P00241_E12 (SEQ ID NO:47), P00245_D06 (SEQ ID NO:48), P00246_D12 (SEQ ID NO:49), P00247_A04 (SEQ ID NO:50), P00248_B04 (SEQ ID NO:52), P00249_F09 (SEQ ID NO:54), P00258_A10 (SEQ ID NO:56), P00262_C10 (SEQ ID NO:58), P00263_G06 (SEQ ID NO:60), P00267_F08 (SEQ ID NO:61), P00269_H08 (SEQ ID NO:62), P00312_C04 (SEQ ID NO:64), P00324_H02 (SEQ ID NO:65), P00628_H02 (SEQ ID NO:66), P00629_C08 (SEQ ID NO:68), P00634_G11 (SEQ ID NO:70), P00641_G11 (SEQ ID NO:71), P00648_E12 (SEQ ID NO:73), P00697_C03 (SEQ ID NO:75);
In another aspect, the invention concerns a composition comprising a polypeptide as hereinabove defined in admixture with a pharmaceutically acceptable carrier. In a specific embodiment, the composition is a pharmaceutical composition, preferably for the treatment of a cardiac, renal or inflanmmatory disease, comprising an effective amount of a polypeptide of the present invention.
In yet another aspect, the invention concerns an antibody specifically binding a polypeptide of the present invention (as hereinabove defined).
In a further aspect, the invention concerns an antagonist or agonist of a polypetide of the present invention.
In a still further aspect, the invention concerns a composition, preferably a pharmaceutical composition, comprising an effective amount of an antibody herein, in admixture with a pharmaceutically acceptable carrier.
The invention further concerns a composition, preferably a pharmaceutical composition, comprising an effective amount of an antagonist or agonist of the present invention, in admixture with a pharmaceutically acceptable carrier.
In a further aspect, the invention concerns a method for the treatment of a cardiac, renal or inflammatory disease, comprising administering to a patient in need an effective amount of a polypeptide of the present invention or an antagonist or agonist thereof.
In a different aspect, the invention concerns a method for the treatment of a cardiac, renal or inflammatory disease, comprising administering to a patient in need an effective amount of a poly- or oligonucleotide of the present invention (as hereinabove defined).
The invention also concerns a method for the treatment of a cardiac, renal or inflammatory disease, comprising administering to a patient in need an effective amount of an antibody specifically binding to a polypeptide of the present invention.
In a further aspect, the invention concerns a method for screening a subject for a cardiac, renal or inflammatory disease characterized by the differential expression of the endogenous homologue of the proteins of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57, 59, 63, 67, 69, 72, 74, or 76 comprising the steps of:
measuring the expression in the subject of the endogenous homologue of the protein of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57, 59, 63, 67, 69, 72, 74, or 76; and
determining the relative expression of such endogenous homologue in the subject compared to its expression in normal subjects, or compared to its expression in the same subject at an earlier stage of development of the cardiac, renal or inflammatory disease. The subject is preferably human and, accordingly, the endogenous protein is a human homologue of the rat proteins of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57, 59, 63, 67, 69, 72, 74, or 76.
In a still further aspect, the invention concerns an array comprising one or more oligonucleotides complementary to reference RNA or DNA encoding a protein of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57, 59, 63, 67, 69, 72, 74, or 76 or another mammalian (e.g. human) homologue thereof, where the reference DNA or RNA sequences are obtained from both a biological sample from a normal subject and a biological sample from a subject exhibiting a cardiac, renal, or inflanmmatory disease, or from biological samples taken at different stages of a cardiac, renal, or inflammatory disease.
In yet another aspect, the invention concerns a method for detecting cardiac, kidney, or inflammatory disease in a human patient comprising the steps of:
providing an array of oligonucleotides at known locations on a substrate, which array comprises oligonucleotides complementary to reference DNA or RNA sequences encoding a human homologue of the proteins of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57, 59, 63, 67, 69, 72, 74, or 76, where the reference DNA or RNA sequences are obtained from both a biological sample from a normal patient and a biological sample from a patient potentially exhibiting cardiac, renal, or inflammatory disease, or from a patient exhibiting cardiac, renal, or inflammatory disease, taken at different stages of such disease (jointly referred to as xe2x80x9cthe test patientxe2x80x9d);
exposing the array, under hybridization conditions, to a first sample of cDNA probes constructed from mRNA obtained from a biological sample from a corresponding biological sample of a normal patient or from a test patient at a certain stage of the disease;
exposing the array, under hybridization conditions, to a second sample of cDNA probes constructed from MRNA obtained from a biological sample obtained from the test patient (if the first sample was taken at a certain stage of the disease, the second sample is taken at a different stage of the disease);
quantifying any hybridization between the first sample of cDNA probes and the second sample of cDNA probes with the oligonucleotide probes on the array; and
determining the relative expression of genes encoding the human homologue of the protein of SEQ ID NO: 2 in the biological samples from the normal patient and the test patient, or in the biological samples taken from the test patient at different stages of the disease.
The invention further concerns a diagnostic kit comprising an array herein (as defined above) for detecting and diagnosing a disease, specifically cardiac, kidney or inflammatory disease. This kit may comprise control oligonucleotide probes, PCR reagents and detectable labels. In addition, this kit may comprise biological samples taken from human subjects, said samples comprising blood or tissue, preferably cardiac tissue, more preferably left ventricle cells. Such diagnostic kits may also comprise antibodies (including poly- and monoclonal antibodies) to a polypeptide of the present invention, including the polypeptide of SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 33, 35, 37, 39, 41, 43, 46, 51, 53, 55, 57, 59, 63, 67, 69, 72, 74, or 76 and further mammalian (e.g. human) homologues thereof.